Existing proteomics technologies can provide a wealth of information about the biochemistry operating in cells. And systems biology tools are being developed to analyze and model this data. However, they fail to address the fundamental questions of how the spatial organization of molecules in cells is established and how it is utilized to control cell function. To answer these, we will need new tools and new theoretical frameworks that specifically include consideration of cell morphology and dynamic spatial molecular distributions. This proposal aims to establish a Technology Center for Networks and Pathways (TCNP) that will integrate microscope technologies for making quantitative in vivo live cell measurements with new physical formulations and computational tools that will produce spatially realistic quantitative models of intracellular dynamics. The model predictions will then be validated with new measurements as well as novel intracellular manipulation technologies also to be developed in our proposed TCNP. These new technologies will be developed and disseminated by the Center for Cell Analysis and Modeling (CCAM) at the University of Connecticut Health Center (UCHC). The technology will be disseminated throughout the research community via training programs, web-based instructional material, a repository of molecular probes and a database of data and models. The proposed work builds on a firm foundation. CCAM is the home of the Virtual Cell, a computational environment for cell biological modeling, and also hosts a variety of projects in biophotonics and live cell microscopic imaging methods as well as a state-of-the-art user facility for nonlinear, confocal, and widefield microscopy. CCAM is the scientific home of an extraordinary confluence of expertise in physics, chemistry, software engineering and experimental cell biology that is unique for a medical school and is ideal for the concerted multi-pronged effort that is planned for the TCNP.